LOCUS       sequence-245798-        7719 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of recombinant P. knowlesi protein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7719)
  AUTHORS   Muller-Sienerth N, Shilts J, Kadir KA, Yman V, Homann MV, Asghar M, 
            Ngasala B, Singh B, Farnert A, Wright GJ
  TITLE     A panel of recombinant proteins from human-infective Plasmodium 
            species for serological surveillance.
  JOURNAL   Malar J. 2020 Jan 17;19(1):31. doi: 10.1186/s12936-020-3111-5.
  PUBMED    31952523
REFERENCE   2  (bases 1 to 7719)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Malar J."; 
            date: "2020-01-17"; volume: "19(1):31. doi"; pages: " 
            10.1186/s12936-020-3111-5"
COMMENT     Sequence Label: Pk_CyRPA-Cd4-bio-His
FEATURES             Location/Qualifiers
     source          1..7719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        7..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..590
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     587..611
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1092..1111
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1134..1143
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2883..2900
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2952..3007
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3006..3025)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5243..5261)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5329..5433
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5434..5502,5503..6294)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5652..5671)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6465..7053
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6954..6973
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7207..7224
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7341..7362
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7377..7382,7383..7400,7401..7407)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7415..7431
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     7420..7442
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7439..7455
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7439..7455
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      241 tacatcaagt gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacacc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
      601 accgtcagat cctcactctc ttccgcatcg ctgtctgcga gggccagctg ttgggctcgc
      661 ggttgaggac aaactcttcg cggtctttcc agtactcttg gatcggaaac ccgtcggcct
      721 ccgaacggta ctccgccacc gagggacctg agcgagtccg catcgaccgg atcggaaaac
      781 ctctcgagaa aggcgtctaa ccagtcacag tcgcaaggta ggctgagcac cgtggcgggc
      841 ggcagcgggt ggcggtcggg gttgtttctg gcggaggtgc tgctgatgat gtaattaaag
      901 taggcggtct tgagacggcg gatggtcgag gtgaggtgtg gcaggcttga gatccagctg
      961 ttggggtgag tactccctct caaaagcggg cattacttct gcgctaagat tgtcagtttc
     1021 caaaaacgag gaggatttga tattcacctg gcccgatctg gccatacact tgagtgacaa
     1081 tgacatccac tttgcctttc tctccacagg tgtccactcc caggtccaag tttgccacca
     1141 tggagtttca gacccaggta ctcatgtccc tgctgctctg catgtctggt gcggccgcca
     1201 acgaggaaag caagcaggtc atcatcctga acgacgagat caccaccatc accagccccg
     1261 tgcactgtat cgccgacacc tacttcatct tccggaacga gctgtacaag atctgcatcc
     1321 agcacgtgaa caagggccgg accgagatcc acgtgatcgt gcagaagaaa gccaagaaca
     1381 agtgggagac aaagcagaag ctgttcgagg acaagatgtg gttccatctg cccttcgtgt
     1441 tcaacttcgt ccagaatgac gagattatca tcctcgtgtg ccggtacaag ggcatgacca
     1501 agggcgaggg cgtggcctgc gatagatggt ctagcaccac cggcaccaac tacaacaagg
     1561 gcaacatcaa catcgacgcc caggccctga ccaagatgaa cctggacagc tacgccagct
     1621 tccccatccc catcaaggac aaggccatca tccacatctg cggcgtgcac agctacgagt
     1681 accagaacgt gaaccagaac aacttcatca gctgcctggc cagcgaggat aagggcacca
     1741 cctggggcga catcaagatt cacatctact acgaccagtt ccaggaaggc gtcccctact
     1801 tctacctgcg gcccctggtg ttcaacgacg agttcggctt ttacctgtac agccggatca
     1861 gcagcaacaa cgccgacaga ggcggcaagt acatgaagtg catcctgaat cccgccaaca
     1921 gccggaacaa agagtacacc ttcaagtgca ccaacgtcaa cctgatcaaa gaggacaagt
     1981 ccctgcagaa tatcgccaag ctgaacggct actacgtgac ctcctacgcc aagaagaaca
     2041 atttcaacga gtgctacctg tactacaccg agaacaacgc catcgtggtc aagcccaagg
     2101 tgcagaacta cgagctgaat ggctgttacg gcggcagctt cgtgaagttc aacgaggcca
     2161 aggccctgtt catctacagc accggccacg gcgtgcagaa catccacacc ctgcactacg
     2221 ccagatacga aggcgcgccg tcgacctcca tcacggccta taagagtgag ggggagtcag
     2281 cggagttctc cttcccactc aaccttggag aggaaagcct gcagggagag ttgagatgga
     2341 aggcagagaa ggctccttct tcccagtcct ggatcacctt ctccctaaag aaccaaaagg
     2401 tgtctgtgca gaagtctact agcaacccca agttccagct gtccgaaacg ctcccactca
     2461 cccttcagat accccaggtc tcccttcagt ttgctggttc tggcaacctg accctgactc
     2521 tggacagagg gatactgtat caggaagtga acctggtggt gatgaaagtg actcagcccg
     2581 acagcaacac tttgacctgt gaggtgatgg gacccacctc acccaagatg agactgatct
     2641 tgaagcagga gaatcaggag gccagggtct ccaggcagga gaaagtgatt caagtgcagg
     2701 cccctgaagc aggggtgtgg caatgtctac tgagtgaagg tgaagaggtc aagatggact
     2761 ccaagatcca ggttttatcc aaagggttga attccggcag cctgcaccac atcctggacg
     2821 cccagaagat gctgtggaac cacagggaca ggaacctgcc tcctctggcc cctctgggcc
     2881 cccaccacca tcaccaccat tgagtctaga cacgtgatta attaaggatc ccccgacctc
     2941 gacctctggc taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg
     3001 tctctcactc ggaaggacat atgggagggc aaatcatttg gtcgagatcc ctcggagatc
     3061 tctagctaga ggatcgatcc ccgccccgga cgaactaaac ctgactacga catctctgcc
     3121 ccttcttcgc ggggcagtgc atgtaatccc ttcagttggt tggtacaact tgccaactgg
     3181 gccctgttcc acatgtgaca cgggggggga ccaaacacaa aggggttctc tgactgtagt
     3241 tgacatcctt ataaatggat gtgcacattt gccaacactg agtggctttc atcctggagc
     3301 agactttgca gtctgtggac tgcaacacaa cattgccttt atgtgtaact cttggctgaa
     3361 gctcttacac caatgctggg ggacatgtac ctcccagggg cccaggaaga ctacgggagg
     3421 ctacaccaac gtcaatcaga ggggcctgtg tagctaccga taagcggacc ctcaagaggg
     3481 cattagcaat agtgtttata aggccccctt gttaacccta aacgggtagc atatgcttcc
     3541 cgggtagtag tatatactat ccagactaac cctaattcaa tagcatatgt tacccaacgg
     3601 gaagcatatg ctatcgaatt agggttagta aaagggtcct aaggaacagc gatatctccc
     3661 accccatgag ctgtcacggt tttatttaca tggggtcagg attccacgag ggtagtgaac
     3721 cattttagtc acaagggcag tggctgaaga tcaaggagcg ggcagtgaac tctcctgaat
     3781 cttcgcctgc ttcttcattc tccttcgttt agctaataga ataactgctg agttgtgaac
     3841 agtaaggtgt atgtgaggtg ctcgaaaaca aggtttcagg tgacgccccc agaataaaat
     3901 ttggacgggg ggttcagtgg tggcattgtg ctatgacacc aatataaccc tcacaaaccc
     3961 cttgggcaat aaatactagt gtaggaatga aacattctga atatctttaa caatagaaat
     4021 ccatggggtg gggacaagcc gtaaagactg gatgtccatc tcacacgaat ttatggctat
     4081 gggcaacaca taatcctagt gcaatatgat actggggtta ttaagatgtg tcccaggcag
     4141 ggaccaagac aggtgaacca tgttgttaca ctctatttgt aacaagggga aagagagtgg
     4201 acgccgacag cagcggactc cactggttgt ctctaacacc cccgaaaatt aaacggggct
     4261 ccacgccaat ggggcccata aacaaagaca agtggccact cttttttttg aaattgtgga
     4321 gtgggggcac gcgtcagccc ccacacgccg ccctgcggtt ttggactgta aaataagggt
     4381 gtaataactt ggctgattgt aaccccgcta accactgcgg tcaaaccact tgcccacaaa
     4441 accactaatg gcaccccggg gaatacctgc ataagtaggt gggcgggcca agataggggc
     4501 gcgattgctg cgatctggag gacaaattac acacacttgc gcctgagcgc caagcacagg
     4561 gttgttggtc ctcatattca cgaggtcgct gagagcacgg tgggctaatg ttgccatggg
     4621 tagcatatac tacccaaata tctggatagc atatgctatc ctaatctata tctgggtagc
     4681 ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt
     4741 atatgctatc ctaatttata tctgggtagc ataggctatc ctaatctata tctgggtagc
     4801 atatgctatc ctaatctata tctgggtagt atatgctatc ctaatctgta tccgggtagc
     4861 atatgctatc ctaatagaga ttagggtagt atatgctatc ctaatttata tctgggtagc
     4921 atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg ggtagcatat
     4981 gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat
     5041 gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg ggtagcatag
     5101 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat
     5161 gctatcctaa tctgtatccg ggtagcatat gctatcctca tgataagctg tcaaacatga
     5221 gaattttctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     5281 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     5341 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5401 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5461 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5521 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     5581 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     5641 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc
     5701 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     5761 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5821 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5881 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5941 gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc
     6001 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     6061 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     6121 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     6181 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     6241 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     6301 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6361 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6421 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6481 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6541 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6601 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6661 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6721 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6781 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6841 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6901 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6961 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     7021 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     7081 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     7141 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     7201 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     7261 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     7321 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     7381 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     7441 ggaaacagct atgaccatga ttacgccaag ctctagctag aggtcgacca attctcatgt
     7501 ttgacagctt atcatcgcag atccgggcaa cgttgttgcc attgctgcag gcgcagaact
     7561 ggtaggtatg gaagatctat acattgaatc aatattggca attagccata ttagtcattg
     7621 gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat ctatatcata
     7681 atatgtacat ttatattggc tcatgtccaa tatgaccgc
//
